Presence of Different Bacterial Species in Thermal Sources and Novelty in Their Industrial Enzyme Productions

Albayrak, Seyda; Genç, Berna; Özkan, Hakan; Taşkın, Mesut; Adıgüzel, Ahmet

doi:10.22207/jpam.13.3.08

Cited by 8 publications

(3 citation statements)

References 48 publications

(43 reference statements)

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“…19 The amplified Journal of Pure and Applied Microbiology polymerase chain reaction fragments were cloned into E. coli JM101 strain with the pGEM-T Easy Cloning Vector (Promega, Southampton, UK) according to the manufacturer instructions. 20,21 The sequences were compared with the other standard bacteria in EzTaxon, and the similarity rate was detected. Then, the GenBank accession number of the P18 was received.…”

Section: Isolation and Molecular Characterization Of P18 From Hot Spr...mentioning

confidence: 99%

Cloning and Production of Protease Enzyme from Aeribacillus pallidus P18 Strain

Saadati,

Baltaci,

Adiguzel

et al. 2024

J. Pure Appl. Microbiol.

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Proteins are essential for the proper functioning of cells. The techniques of cloning and protein production have facilitated the advancement of various fields and the creation of specific proteins for industrial and therapeutic uses. The bacterium Aeribacillus pallidus, which is able to survive in extreme conditions, is being studied with a view to identifying its robust enzymes. The objective of this study was to clone the protease gene from the A. pallidus P18 strain into the SUMO vector and produce recombinant protein in Escherichia coli BL21 for protein production. The protease enzyme gene from the A. pallidus P18 strain was isolated and amplified by using PCR. The PCR product was transferred into the SUMO expression vector and amplified in One Shot® Mach1TM-T1R bacteria, followed by colony PCR. Plasmid isolation was performed after positive colony selection. Gene integration was confirmed by cross-PCR using the gene forward, and vector reverse primers. For expression, the plasmid was transferred to E. coli BL21 cells. Two cultures were induced with different IPTG concentrations (0.5 mM and 1 mM) to optimize protein production. Bacterial cells were lysed, and SDS-PAGE analysis was conducted. Purification involved cell lysate preparation and purification using a ProbondTM column. SDS-PAGE and Coomassie Brilliant Blue G-250 staining confirmed successful purification. The results of this study indicate that the optimal product for protein production is that derived from a culture induced with 1 mM IPTG. Upon completion of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) procedure, the weight mass of the produced protein was determined to be 37 kDa, as indicated by the result of the gel stained with Coomassie brilliant blue G-250. This research successfully cloned the protease enzyme gene from the A. pallidus P18 strain using the pET-SUMO vector, performed purification and achieved the targeted result of protein production.

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Section: Isolation and Molecular Characterization Of P18 From Hot Spr...mentioning

confidence: 99%

Cloning and Production of Protease Enzyme from Aeribacillus pallidus P18 Strain

Saadati,

Baltaci,

Adiguzel

et al. 2024

J. Pure Appl. Microbiol.

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“…The bacterial isolates were inoculated on tributyrin agar (23 g/L) medium which contained 1% tributyrin (glycerol tributyrate) and incubated at 15°C for 3-5 days. The strains with transparent and the highest zone formation (lipolytic activity) were determined as lipase producers 16 . Amylase Amylase assay was performed at 15°C for 3-5 days in a medium which contained nutrient agar (20 g/L), soluble starch (5 g/L) and agar (15 g/L).…”

Section: Lipasementioning

confidence: 99%

Proteolytic, Lipolytic and Amylolytic Bacteria Reservoir of Turkey; Cold-Adaptive Bacteria in Detergent Industry

Muslu¹,

Genç²,

Mahmut³

et al. 2020

J. Pure Appl. Microbiol.

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Enzymes which would be active in cold conditions can be used in a wide range of fields from molecular biology to detergent industry due to their low processing capacity and high activity. In this study, sixty cold-adapted bacteria were isolated from water and sludge samples collected from Erzurum and Van provinces. Identification of eight isolates by molecular [(GTG) 5 -PCR and 16S rRNA sequencing] techniques and tests for temperature (4-35°C), pH (3-11) and salt (2-15% (w/v) requirements were performed. These bacteria were belonging to Pseudomonas chlororaphis subsp. aureofaciens (SM 01 1 A ), Psychrobacter faecalis (SM 01 2 D ), Rahnella aquatilis (SM 01 5 A ), Shewanella putrefaciens (SM 01 8 A ), Pseudomonas lactis (SM 01 10 A ), Flavobacterium chryseum (SM 01 12 E ), Exiguobacterium mexicanum (SM 01 17 A ) and Glutamicibacter arilaitensis (SM 01 18 A). The physicochemical requirements for all isolates ranged between 4-25°C, pH 5-7 and 2-15% salt (NaCl) concentration. However, E. mexicanum did not require salt in growth medium. All bacteria were evaluated for protease, lipase and amylase enzymes and all were found to be multiple enzyme producers. The eight isolates were identified from the resources of Turkey, for the first time and enzyme production abilities of some isolates to produce enzymes were declared. The originating of the producers of these enzymes from Turkey shows that Turkey has a remarkable reservoir for cold-adaptive microorganisms and these microorganisms will make important contributions to the detergent industry worldwide.

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“…Afterwards, test isolates were subjected to rep-PCR [(GTG) 5 -PCR] with the special primer of (GTG) 5 elements to obtain genomic fingerprinting. The 16S rRNA gene regions of the isolates which would be considered to be different according to the rep-PCR analysis were amplified with universal primers by PCR and were subjected to the cloning procedure [13][14][15][16][17] .…”

Section: Isolation and Molecular Characterization Of Lactic Acid Bacteriamentioning

confidence: 99%

Bacteriocin Producing Bacteria Isolated from Turkish Traditional Sausage Samples

Erkaya¹,

Genç²,

Akbulut³

et al. 2020

J Pure Appl Microbiol

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In this study, traditional sausage samples from different provinces of Turkey (Gaziantep, Antalya, erzurum and Kahramanmaras) were obtained and one hundred three isolates were collected. Using the (GTG) 5 -PCR genomic fingerprint analysis method, seven of them were observed to be different and conventional tests of these isolates were performed. Molecular identification of two isolates carrying the bacteriocin gene and having antimicrobial activity by agar disc diffusion method was performed by 16S rRNA sequence analysis. As a result, the seven isolates were identified as Aerococcus urinaeequi (EK1), Streptococcus salivarius (EK2), Leuconostoc mesenteroides (EK3), Macrococcus caseolyticus (EK4), Lactococcus garvieae (EK5), Staphylococcus saprophyticus (EK6) and Lactobacillus sakei (EK7). Among these strains, it has been determined that Ln. mesenteroides and L. sakei carried the mecentericin and sacacin genes. When antimicrobial activity against different strains was examined, inhibition formations of Ln. mesenteroides and L. sakei on Enterococcus faecalis, Shigella dysenteriae and Escherichia coli O157: H7 were observed.

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Presence of Different Bacterial Species in Thermal Sources and Novelty in Their Industrial Enzyme Productions

Cited by 8 publications

References 48 publications

Cloning and Production of Protease Enzyme from Aeribacillus pallidus P18 Strain

Cloning and Production of Protease Enzyme from Aeribacillus pallidus P18 Strain

Proteolytic, Lipolytic and Amylolytic Bacteria Reservoir of Turkey; Cold-Adaptive Bacteria in Detergent Industry

Bacteriocin Producing Bacteria Isolated from Turkish Traditional Sausage Samples

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